Quantitative Analysis of Prenylated RhoA Interaction with Its Chaperone, RhoGDI

Z Tnimov, Z Guo, Y Gambin, Nguyen UTT, Wu YW, Daniel Abankwa, A Stigter, BM Collins, H Waldmann, RS Goody, K Alexandrov

Tutkimustuotos: LehtiartikkeliArtikkeliTieteellinenvertaisarvioitu

29 Sitaatiot (Scopus)

Abstrakti

Small GTPases of the Rho family regulate cytoskeleton remodeling, cell polarity, and transcription, as well as the cell cycle, in eukaryotic cells. Membrane delivery and recycling of the Rho GTPases is mediated by Rho GDP dissociation inhibitor (RhoGDI), which forms a stable complex with prenylated Rho GTPases. We analyzed the interaction of RhoGDI with the active and inactive forms of prenylated and unprenylated RhoA. We demonstrate that RhoGDI binds the prenylated form of RhoA center dot GDP with unexpectedly high affinity (K-d = 5 pM). The very long half-life of the complex is reduced 25-fold on RhoA activation, with a concomitant reduction in affinity (K-d = 3 nM). The 2.8-angstrom structure of the RhoA center dot guanosine 5'-[beta,gamma-imido] triphosphate (GMPPNP)center dot RhoGDI complex demonstrated that complex formation forces the activated RhoA into a GDP-bound conformation in the absence of nucleotide hydrolysis. We demonstrate that membrane extraction of Rho GTPase by RhoGDI is a thermodynamically favored passive process that operates through a series of progressively tighter intermediates, much like the one that is mediated by RabGDI.
AlkuperäiskieliEi tiedossa
Sivut26549–26562
Sivumäärä14
JulkaisuJournal of Biological Chemistry
Vuosikerta287
Numero32
DOI - pysyväislinkit
TilaJulkaistu - 2012
OKM-julkaisutyyppiA1 Julkaistu artikkeli, soviteltu

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